INVESTIGATING NEUTRALIZING ANTIBODY INDUCED CONFORMATIONAL CHANGES AND MATURATIO

这个子项目是许多利用资源的研究子项目之一 由NIH/NCRR资助的中心拨款提供。子项目的主要支持 而子项目的主要调查员可能是由其他来源提供的， 包括其它NIH来源。 列出的子项目总成本可能 代表子项目使用的中心基础设施的估计数量， 而不是由NCRR赠款提供给子项目或子项目工作人员的直接资金。 已知结构的对照蛋白和几种gp 140三聚体形式的先前SAXS结果表明，SAXS是我们掌握的一种易于处理的分析方法。所有样品都可以在短时间内制备，用于SAXS数据收集，目前正在进行常规生产，以支持其他研究。通过SEC同时进行静态/动态光散射以及多种形式的定量/定性Ab结合试验监测蛋白质单分散性。 我们期望获得不同gp 140-NAb复合物的整体形状和大小信息，如果可能的话，生成包膜尖峰的功能形式的模型，并通过冷冻电镜结构的Env尖峰和晶体结构的gp 120和gp 41进行验证。

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. Primary support for the subproject and the subproject's principal investigator may have been provided by other sources, including other NIH sources. The Total Cost listed for the subproject likely represents the estimated amount of Center infrastructure utilized by the subproject, not direct funding provided by the NCRR grant to the subproject or subproject staff. Prior SAXS results with control proteins of known structure and several gp140 trimer forms show that SAXS is a tractable method of analysis in our hands. All samples can be prepared on short notice for SAXS data collection and are currently in routine production to support other studies. Protein monodispersity is monitored by SEC with simultaneous static/dynamic light scattering as well as quantitative/qualitative Ab binding assays in a variety of formats. We expect to obtain overall shape and size information of different gp140-NAb complexes, generate models for the functional form of envelope spike if possible, and verify by cryo-EM structures of Env spikes and crystal structures of gp120 and gp41.